LOCUS pEN-L4-1-R1 2930 bp DNA circular 06-SEP-2022 DEFINITION . ACCESSION urn.local...oy-dt3gjrs KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/ VNTDATE|459284646| VNTDBDATE|459284646| LSOWNER| VNTNAME|pEN-L4-1-R1,0| VNTAUTHORNAME|Demo User| Vector_NTI_Display_Data_(Do_Not_Edit!) 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(CObjectList #54=(CGroupPar (CParagraph 0 (10 #6# 0) 1 2 2 0 180) (CObjectList #55=(CLinePar (CParagraph 0 (1 #6# 1) 1 2 2 0 191) " N1: 152 " 1))))) "EcoRI: 1 site") 1 5 "GAATTC" "CTTAAG") #56=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 40826984 (8 0) 1 1 1 0 178) (CObjectList #57=(CGroupPar (CParagraph 0 (10 #7# 0) 1 2 2 0 180) (CObjectList #58=(CLinePar (CParagraph 0 (1 #7# 1) 1 2 2 0 191) " N1: 186 " 1))))) "EcoRV: 1 site") 3 3 "GATATC" "CTATAG") #59=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 40858000 (8 0) 1 1 1 0 178) (CObjectList)) "HindIII: 0 sites") 1 5 "AAGCTT" "TTCGAA") #60=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 41628280 (8 0) 1 1 1 0 178) (CObjectList #61=(CGroupPar (CParagraph 0 (10 #9# 0) 1 2 2 0 180) (CObjectList #62=(CLinePar (CParagraph 0 (1 #9# 1) 1 2 2 0 191) " N1: 2530 " 1))))) "NcoI: 1 site") 1 5 "CCATGG" "GGTACC") #63=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 41627160 (8 0) 1 1 1 0 178) (CObjectList #64=(CGroupPar (CParagraph 0 (10 #10# 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FEATURES Location/Qualifiers attL4 1..97 /Color="#00cccc" /created_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attL4" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL4" misc_feature 99^100 /Original_Bases="CTGCA" /label="CTGCA" /note="Geneious type: Editing History Deletion" promoter 106..378 /created_by="thjac" /modified_by="mamee" /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /X-Transferred_From="pNOS" /X-Transferred_Similarity="100.00%" /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; pNOS" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="Basta" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; pNOS (Basta)" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; pNOS" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; pNOS (Hygromycin)" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; pNOS (Kanamycin)" /Alternative_Match="99.64%; Adjusted-Interval-0:-6/0; pNOS" /label="pNOS" misc_feature 203 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" attR1 385..541 /created_by="mamee" /Color="#00cccc" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attR1" /Transferred_Similarity="99.36%" /Primary_Match="99.36%; Adjusted-Interval-0:0/0; attR1" misc_feature 511 /Original_Bases="T" /label="T" /note="Geneious type: Editing History Replacement" primer_bind 568..587 /%GC="60.0" /Hairpin_Tm="None" /Self_Dimer_Tm="8.2" /Tm="59.3" /annotation_group="6052266140032900d7ed8dbf-ddf2-4504-acc5 -13d826d327d1" /created_by="primer3" /Sequence="GGCCGCGATATCCCCTATAG" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-1-R1" /label="P40" primer complement(601..621) /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCDB-Cas9 - GWM13R" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13R (pCDB-Cas9 - GWM13R)" /label="GWM13R" promoter 682..732 /created_by="chgai" /X-Transferred_From="pCG034_pKan_Kan-CBE" /X-Transferred_Similarity="100.00%" /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG034_pKan_Kan-CBE)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG040_ABE-Nuc-NLS_Kan -reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG043_Kan-ABE-Dest_co ntrol_ccdB)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG046_ABE-LbCas12a-NL S_Kan_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG005_pGGD-pKan-KanR- E)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG006_pGGA-Kan_promot er-B)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG035_pKan_Kan)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG039_ABE-N7-NLS_Kan- reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG041_ABE-SV40-NLS_Ka n-reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG054_Kan_CBE_control )" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="KanR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (KanR)" /label="kan" misc_feature 733..1542 /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="KanR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; KanR" /label="KanR" /note="Geneious type: Bacterial Selection" primer_bind 1392..1411 /%GC="60.0" /Hairpin_Tm="40.0" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6052266197937900564efd27-a78d-4910-9c6b -b41817c1cb7c" /created_by="primer3" /Sequence="CGCAGACCGATACCAGGATC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-1-R1" /label="P42" rep_origin 1689..2277 /created_by="thjac" /modified_by="mamee" /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ColE1/pMB1/pBR322/pUC Ori (high-copy-number ColE1/pMB1/pBR322/pUC origin of replication)" /label="ColE1/pMB1/pBR322/pUC Ori" misc_feature 1836 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" misc_feature 2081..2082 /Original_Bases="AT" /label="AT" /note="Geneious type: Editing History Replacement" primer_bind 2177..2196 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-1-R1" /Sequence="GGGGAAACGCCTGGTATCTT" /Tm="59.7" /%GC="55.0" /Hairpin_Tm="37.3" /Self_Dimer_Tm="None" /modified_by="mamee" /label="P45" primer_bind 2785..2804 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="605226643142200067104fe5-fbd3-487d-a4ab -ada74a6b2d4b" /created_by="primer3" /Sequence="CTTCCGACTGAGCCTTTCGT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L4-1-R1" /label="P46" primer 2876..2896 /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCDB-Cas9 - GWM13F" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13F (pCDB-Cas9 - GWM13F)" /label="GWM13F" misc_feature 2909 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" ORIGIN 1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa 61 tgctttttta taatgccaac tttgtataga aaagttggaa tacatgagaa ttaagggagt 121 cacgttatga cccccgccga tgacgcggga caagccgttt tacgtttgga actgacagaa 181 ccgcaacgtt gaaggagcca ctgagccgcg ggtttctgga gtttaatgag ctaagcacat 241 acgtcagaaa ccattattgc gcgttcaaaa gtcgcctaag gtcactatca gctagcaaat 301 atttcttgtc aaaaatgctc cactgacgtt ccataaattc ccctcggtat ccaattagag 361 tctcatattc actctcaact cgaacaagtt tgtacaaaaa agttgaacga gaaacgtaaa 421 atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact 481 gtaaaacaca acatatgcag tcactatgaa ccaactactt agatggtatt agtgacctgt 541 agaattcgag ctctagagct gcagggcggc cgcgatatcc cctatagtga gtcgtattac 601 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt 661 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat 721 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac 781 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg 841 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa 901 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt 961 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc 1021 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa 1081 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat 1141 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac 1201 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc 1261 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat 1321 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga 1381 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag 1441 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg 1501 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg 1561 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa 1621 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa 1681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 1741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 1801 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 1861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 1921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 1981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 2041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 2101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 2161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 2221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 2281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 2341 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 2401 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 2461 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 2521 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct 2581 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag 2641 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca 2701 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac 2761 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg 2821 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa 2881 acgacggcca gtcttaagct cgggcccgag ttaacgctac catggagctc //